In order to follow the dynamics of single cellular micro-tubules in living cells using time-lapse imaging, expression at high levels of Green-Fluorescent Protein linked to the microtubule associated protein were required. Traditional Green Fluorescent Protein labeling methods induced artifacts such as micro-tubule bundling, mitotic abnormalities, photo bleaching (Heim, et al., 1996) (Cormack et al., 1996)and on/off blinking (Dickson et al, 1997). These labeling induced changes in cell morphology limits the usefulness of traditional Green Fluorescent Protein labeling in vivo.
This invention discloses a method for observing in vivo dynamics and function critical for many cellular processes in living cells using a Green Fluorescent Protein chimeric molecule (GFP-Ensconsin) to provide a non-perturbing label of cellular components.
This invention discloses that the Green Fluorescent Protein chimeric molecule produces a strong stable fluorescence signal useful in labeling individual protein components making them visible for quantitation by fluorescence speckle microscopy and time lapse imaging.